Hybrid varieties dominate the red beet market. The breeding process necessary to produce these cultivars is very difficult and time consuming. The application of in vitro gynogenesis can reduce the time needed to produce the corresponding homozygous pure lines to a few months. Our research team has developed a method to obtain red beet doubled haploid plants by gynogenesis. The best medium for gynogenesis induction is the B5 medium with the addition of 0.5 mg/L IAA, 0.2 mg/L BA, and 322 mg/L putrescine, whereas the best medium for shoot induction from embryos proved to be the MS medium supplemented with 0.1 mg/L NAA, 0.1 mg/L BA, and 0.5 mg/L putrescine. https://www.selleckchem.com/GSK-3.html The shoots obtained were rooted on MS medium containing half the concentration of microelements and 3 mg/L NAA, 160 mg/L putrescine, and 20 g/L sucrose. Ploidy evaluation of gynogenetic plants was performed by flow cytometry and homozygosity or heterozygosity was determined by two isoenzymatic systems PGI and AAT.Doubled haploid technology allows for producing completely homozygous plants in one generation, which is a very efficient and fast method compared to the production of near-homozygous lines by selfing through conventional breeding methods. However, grain legumes are known to be recalcitrant for most of the in vitro approaches to doubled haploidy. In the last years, significant advances have been made with several legume species through in vitro methods. Chickpea is one of the most important legume species. Several reports have documented the successful generation of haploid plants through anther culture. These reports also showed that successful production of chickpea haploids was achieved when time- and labor-consuming physical stresses such as centrifugation and electroporation were applied to anthers as a pretreatment. In this chapter, we present an efficient and simple anther culture protocol for production of chickpea haploid plants using high concentrations of 2,4-D and silver nitrate in the culture medium, but without applying any physical stresses to anthers.Homozygous parental lines are indispensable for commercial hybrid seed production in many ornamental and vegetable crops. The in vitro induction of haploids and doubled haploids (DHs) through gametic embryogenesis is an effective approach for single-step development of complete homozygous lines from heterozygous donor plants. Anther culture is one of the most popular and widely employed techniques for development of haploids. Here we describe the detailed protocol for rapid and successful induction of haploids in Tagetes spp. using in vitro androgenesis approach. In this protocol, we have provided the comprehensive details of various steps of anther culture in marigold right from the growing of donor plants, selection of buds, pretreatment, embryogenesis and regeneration to ploidy analysis, and chromosome doubling for development of DHs.The production of doubled haploid (DH) plants from microspores is an important technique used in plant breeding and basic research. DH technology is a rapid method for developing homozygous lines, which can be used to accelerate crop improvement programs. Haploidy technology can also be used in mutagenesis, transformation, and basic research such as genomic, biochemical, and physiological studies. There is no general protocol that will result in the production of DH in all species, as differences occur among species and among genotypes within a species in terms of embryogenic response. Here we describe methodology for developing doubled haploids in cow cockle (Saponaria vaccaria L.).African violet (Saintpaulia ionantha) is an herbaceous perennial of the Gesneriaceae family. Because almost all the cultivars are heterozygous, pure lines are useful for both classical and new breeding approaches. A shortcut to obtain purebred lines involves the production of doubled haploid strains produced from anther-derived haploids. In this chapter, a protocol for culturing African violet anthers is described in detail.Borage (Borago officinalis L.) is a crop with different culinary, pharmaceutical, and industrial properties. Besides, it is one of the best known sources of gamma linolenic acid (GLA). However, the variability in the levels of such active compounds, obtained from wild borage, may result in conflicting clinical trial reports, which may likely decrease the optimal efficiency of the product. On the other hand, this important medicinal plant has a multifactorial self-incompatibility system, which makes self-pollination ineffective and results in a limited production of pure (homozygous) lines for breeding programs. To avoid the limitations of self-incompatibility and also producing uniform lines useful as parents for F1 hybrid production, or as starting materials to develop new varieties with high and homogenous levels of medicinal compounds, androgenic doubled haploid (DH) lines produced by anther culture have the potential to speed up the process of producing homozygous lines for breeding program of this medicinal species. In the present chapter, a protocol for production of haploid plants in borage by in vitro anther culture is described.The obstacles to breeding programs in Jatropha are the long reproductive cycle with a juvenile phase that lasts several months, the highly heterozygous nature of the genome, the large canopy size, and self-incompatibility that is a long-term process which requires multiple cycles of self-pollination to achieve complete homozygosity. In vitro plant tissue culture-based tools such as haploids and doubled haploid techniques can increase the selection efficiency, resulting into selection of superior plants with complete homozygosity in one generation. It bypasses the complications of greenhouse field evaluation or off-season generation advancement, which takes about 8-10 generations in traditional breeding with the time line of 10-12 years. The haploids have in fact a single set of chromosomes, which undergoes duplication spontaneously during in vitro culture conditions, and are further converted into doubled haploid plants. This represents a major biotechnological tool to accelerate plant breeding. Here, we have established a reproducible, unique anther culture protocol in Jatropha curcas to develop haploid and doubled haploid plants.